<p>Since plants can not escape their predators by walking, they use some other defense systems, like induction or repression of defense genes. A microarray experiment performed with barley attacked by the bird cherry-oat aphid (Rhopalosiphum padi), led to the hypothesis that contig 16360 (similar to ser/thr kinases) could be linked with the resistance of barley against R. padi, and contig 6519 (similar to WIR 1A) with the susceptibility. Time course experiments showed that contig16360 and AJ250283 (similar to BCI-4) are almost induced in the same way, each, by two different aphids (R. padi and Metopolophium dirhodum). Genomic PCR was used to test the hypothesis that when plants have the gene for contig 16360, they are more likely to be resistant against aphid attack, and when plants have the gene for contig 6519, they are more likely to be susceptible. This test was performed with 69 barley lines: wild, commercial or breeding lines. Results were that the presence of WIR 1A gene has no correlation with the susceptibility, while presence of ser/thr kinase seems to be correlated with resistance.</p>

Since plants can not escape their predators by walking, they use some other defense systems, like induction or repression of defense genes. A microarray experiment performed with barley attacked by the bird cherry-oat aphid (Rhopalosiphum padi), led to the hypothesis that contig 16360 (similar to ser/thr kinases) could be linked with the resistance of barley against R. padi, and contig 6519 (similar to WIR 1A) with the susceptibility. Time course experiments showed that contig16360 and AJ250283 (similar to BCI-4) are almost induced in the same way, each, by two different aphids (R. padi and Metopolophium dirhodum). Genomic PCR was used to test the hypothesis that when plants have the gene for contig 16360, they are more likely to be resistant against aphid attack, and when plants have the gene for contig 6519, they are more likely to be susceptible. This test was performed with 69 barley lines: wild, commercial or breeding lines. Results were that the presence of WIR 1A gene has no correlation with the susceptibility, while presence of ser/thr kinase seems to be correlated with resistance.

The broad aim of the work described in this thesis was to use the arbuscular mycorrhizal (AM) defective rmc tomato to explore the development and function of different types of fungus-plant interfaces (phenotypes) and to characterize the cellular modifications preceding colonization of rmc by a range of different AM fungi. Three main patterns of colonization with rmc have been described: 1) Pen- phenotype in which the AM fungus is restricted to the root surface with several attempts to penetrate the epidermal cells without success; 2) Coiphenotype where AM fungi penetrate the epidermis but cannot develop cortical colonization; and 3) Myc+ phenotype (with G. intraradices WFVAM23), where the AM fungus penetrates the cortex and forms a “normal” colonization after a delayed penetration of the epidermal cells (Review of literature). Little is known about cellular interactions, nutrient transfer or the ability of the fungi to complete their life cycles in the different phenotypes. These aspects were the main foci of this work. In addition further fungal isolates were screened to asses their ability to colonize rmc. The first experiments involved compartmented pots to follow the fungal life cycle, production of external mycelium and spores in the different rmc phenotypes (Chapter 3). The results showed that in the Pen- and Coiphenotypes, AM fungi are unable to form spores to complete the life cycle. However, in the Coi-phenotype, the fungus remained alive up to week 18, suggesting that some C transfer occurred. The fungus forming the Myc+ phenotype, G. intraradices WFVAM23, was able to produce spores, although they were significantly smaller than those produced with the wild-type tomato. The results suggested that arbuscules are essential for completion of the fungal life cycle. Labeled 32P was used to determine whether arbuscules are also essential for P transfer (Chapter 4). A compartmented pot system was used in which only fungal hyphae but not roots could obtain 32P. 32P was found in the shoots of rmc inoculated with S. calospora (Coi- phenotype), indicating that interfaces other than arbuscules can be involved in transfer of P. A nurse pot system was used to obtain synchronized colonization to determine how long AM fungi stay alive during the interactions with rmc and to elucidate the cellular modifications preceding colonization of rmc by a range of different AM fungi (Chapter 5). The results showed that rmc did attract the AM fungi, that the plant nucleus moved to the middle of the plant cell only after fungal penetration of plant roots and that callose deposition in rmc was not involved in blocking the AM fungi. Fourteen AM fungi with different taxonomic affiliations and fourteen different G. intraradices isolates were screened to try to relate phylogeny of AM fungi with phenotypes in rmc (Chapter 6). There were a large number of interactions, depending on the inoculated AM fungi, and although there were some similarities in the rmc phenotypes within phylogenetic groups, there was no clear relationship between phylogeny and development of interactions with rmc. This study showed the following. 1) Arbuscules/arbusculate coils are necessary for the completion of the AM fungal cycle. However, intraradical hyphae also participate in transfer of both P and C as demonstrated with the Coi- phenotype. 2) rmc clearly attracted AM fungi and the fungi stay alive and induce plant cellular responses such as nuclear movement only after penetrating rmc roots. 3) Plant defense responses such as callose deposition are not involved in blocking AM fungi in rmc; and 4) there was no relationship between the phenotypes described in rmc and phylogeny of the Glomeromycota. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1292816 / Thesis(Ph.D.)-- School of Earth and Environmental Sciences, 2007.

Ce travail s’inscrit dans l’étude de l’incidence de sidérophores sur la physiologie de la plante. Il décrit plus précisément, à l’échelle phénotypique et moléculaire, l’impact de la pyoverdine produite par la souche bactérienne bénéfique Pseudomonas fluorescens C7R12 sur la croissance, la réponse immunitaire et l’homéostasie du fer chez Arabidopsis thaliana. Le lien fonctionnel entre immunité et homéostasie du fer a été abordé de façon plus spécifique via l’analyse du mode d’action de l’acide β-aminobutyrique (BABA), un potentialisateur des réponses de défense de la plante. En conditions de fer limitantes, afin de pourvoir à la carence, Pseudomonas fluorescens libère la pyoverdine dans le sol sous sa forme non chélatée (apo-pyo). Le complexe fer-pyoverdine (ferri-pyo) est ensuite internalisé par la bactérie. Nous avons vérifié que l’apo-pyo est assimilée par des plantes d’A. thaliana cultivées dans un milieu contenant ou non du fer. De façon remarquable, l’apo-pyo restaure le phénotype de croissance des plantes carencées en fer. Une analyse transcriptomique a révélé que chez ces dernières, l’apo-pyo induit fortement l’expression de gènes associés à la croissance, l’import et la redistribution du fer in planta. En revanche, une répression de l’expression de gènes de défense s’opère. De façon concordante, l’effet promoteur de croissance de l’apo-pyo chez les plantes carencées est strictement dépendant de l’expression des gènes IRT1 et FRO2 codant deux protéines majeures de l’import de fer. De plus, une moindre résistance de ces plantes à Botrytis cinerea a été relevée. L’incidence négative de l’apo-pyo sur les défenses s’accompagne d’une surexpression du facteur de transcription HBI1 jouant un rôle clé dans la régulation de la balance croissance/défense. L’ensemble de ces événements n’a pas été observé chez les plantes cultivées dans un milieu enrichi en fer, démontrant que les effets de l’apo-pyo chez A. thaliana sont conditionnés par le statut en fer de la plante. En parallèle, l’étude du mode d’action du BABA a indiqué que ce potentialisateur de l’immunité est un chélateur très efficace du fer. En conséquence, appliqué à des plantes d’A. thaliana, le BABA déclenche une carence en fer transitoire. Nous avons émis l’hypothèse que cette carence pourrait constituer un signal plaçant la plante en veille défensive. En accord avec cette assomption, les plantes carencées en fer présentent une résistance accrue à B. cinerea et produisent des métabolites secondaires associés aux défenses dont l’accumulation est également induite par le BABA. Ainsi, la carence en fer transitoire occasionnée par le BABA pourrait constituer l’une des composantes de son effet potentialisateur sur l’immunité. En conclusion, ce travail apporte des premiers éléments explicatifs quant à l’incidence de la pyoverdine sur des traits physiologiques de la plante et rapporte un mode d’action orignal du BABA. Plus généralement, il renforce le concept encore naissant de l’existence de régulations croisées entre les voies de signalisation associées à la croissance, l’immunité et l’homéostasie du fer chez les plantes. / Siderophores are strong iron chelators produced by bacteria under iron deficiency conditions. In the present work, we studied the impact of the siderophore pyoverdine, produced by the plant growth promoting rhizobacteria Pseudomonas fluorescens C7R12, on plant physiology from phenotypic to molecular effects with a specific focus on plant growth, immune response and iron homeostasis. Based on our analysis of the mode of action of the non-protein amino acid β-aminobutyric acid (BABA), a priming inducer in plants, we studied more specifically the functional link between iron homeostasis and plant immunity. Under iron deficiency, P. fluorescens excretes the iron free form of pyoverdine (apo-pyo) in the soil. Once chelated with iron (ferri-pyo), the complex is internalized by the bacteria. We demonstrated that Arabidopsis thaliana plants treated by apo-pyo in a medium containing or not iron (Fe 25 or Fe 0) also internalize pyoverdine. Moreover, we observed that under iron deficiency, pyoverdine treated plants did not display the growth reduction induced by iron deficiency. In accordance with this phenotype, a microarray analysis revealed that the expression of genes related to growth and development was induced, as well as genes related to iron uptake and transport in planta. In contrast, the down regulation of the expression of genes related to defense was observed. Correspondingly, we demonstrated that the growth improvement induced by apo-pyo under iron deficiency depends on the expression of IRT1 and FRO2, two major genes involved in iron uptake mechanisms. Of interest, the resistance to Botrytis cinerea conferred by iron deficiency was lost following apo-pyo treatment. The overexpression of the HBI1 transcription factor, known to be involved in the growth-defense tradeoff, can be linked to the above observations. These apo-pyo effects were not observed after treatment of plants under sufficient iron conditions, indicating that in A. thaliana apo-pyo effects are dependent on the plant iron status. In the same time, the analysis of the mode of action of BABA that potentiates plant defense responses demonstrated that BABA is a powerful iron chelator. BABA treatment in A. thaliana triggered a transient iron deficiency response. Based on this assessment, we assume that iron deficiency response and priming of defense may be connected. In accordance with this hypothesis, we showed that plants cultivated under iron deficiency and BABA treated plants both displayed resistance to B. cinerea and produced secondary metabolites associated to defense. Hence, the BABA priming effects on plant defense may be due to the induction of transient iron deficiency. To conclude, this work draws first explications on pyoverdine effects on plant physiology and presents an original mode of action contributing to the priming effects of BABA. In a larger view, this work supports the recent concept of the existence of a cross-regulation between growth, immunity and iron homeostasis in plants.